Rapid mesoscale volumetric imaging of neural activity with synaptic resolution

Lü, Rong; Liang, Yajie; Meng, Guanghan; Zhou, Pengcheng; Svoboda, Karel; Paninski, Liam; Ji, Na

doi:10.1038/s41592-020-0760-9

Cited by 104 publications

(91 citation statements)

References 28 publications

(33 reference statements)

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“…The second order term is used to calculate the GDD, which for our laser system was ~ -590 fs 2 . The third order term is used to calculate the TOD, which for our laser was ~ 22,750 fs 3 . Because the goal was to estimate the amount of TOD in the laser source, we performed these measurements at the laser head [see Supplementary Fig.…”

Section: A Comprehensive Measurement Of Laser Pulse Propertiesmentioning

confidence: 99%

“…The now ubiquitous field of two-photon microscopy has been used to visualize a wide array of biological phenomena in vivo, from imaging neurons in healthy animals [1][2][3] and in disease models such as Alzheimer's 4 , to imaging the movement of fluorescently-labeled viruses invading the lungs 5,6 and the mechanisms of glomerular filtration in the kidneys 7 . Three-photon microscopy builds on the success of two-photon microscopy by using longer excitation wavelengths to reduce light scattering and a three-photon absorption process to achieve better nonlinear confinement in biological tissue, resulting in higher signal to background ratio at larger depths [8][9][10][11] .…”

Section: Introductionmentioning

confidence: 99%

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Improving laser standards for three-photon microscopy

Farinella

Roy

Liu

et al. 2020

Preprint

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SignificanceThree-photon excitation microscopy has double-to-triple the penetration depth in biological tissue over two-photon imaging and thus has the potential to revolutionize the visualization of biological processes in vivo. However, unlike the ‘plug-and-play’ operation and performance of lasers used in two-photon imaging, three-photon microscopy presents new technological challenges that require a closer look at the fidelity of laser pulses.AimWe implemented state-of-the-art pulse measurements and developed new techniques for examining the performance of lasers used in three-photon microscopy. We then demonstrated how these techniques can be used to provide precise measurements of pulse shape, pulse energy and pulse-to-pulse intensity variability, all of which ultimately impact imaging.ApproachWe built inexpensive tools, e.g., a second harmonic generation frequency resolved optical gating (SHG-FROG) device, and a deep-memory diode imaging (DMDI) apparatus, to examine laser pulse fidelity.ResultsFirst, SHG-FROG revealed very large third order dispersion (TOD). This extent of phase distortion prevents the efficient temporal compression of laser pulses to their theoretical limit. Furthermore, TOD cannot be quantified when using a conventional method of obtaining the laser pulse duration, e.g., when using an autocorrelator. Finally, DMDI showed the effectiveness of detecting pulse-to-pulse intensity fluctuations on timescales relevant to three-photon imaging, which were otherwise not captured using conventional instruments and statistics.ConclusionsThe distortion of individual laser pulses caused by TOD poses significant challenges to three-photon imaging by preventing effective compression of laser pulses and decreasing the efficiency of nonlinear excitation. Moreover, an acceptably low pulse-to-pulse amplitude variability should not be assumed. Particularly for low repetition rate laser sources used in three-photon microscopy, pulse-to-pulse variability also degrades image quality. If three-photon imaging is to become mainstream, our diagnostics may be used by laser manufacturers to improve system design and by end-users to validate the performance of their current and future imaging systems.

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Section: A Comprehensive Measurement Of Laser Pulse Propertiesmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Improving laser standards for three-photon microscopy

Farinella

Roy

Liu

et al. 2020

Preprint

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“…Recent advances in electrophysiology and calcium imaging have extended their spatial scale (Lu et al, 2020;Musall et al, 2019;Sofroniew et al, 2016;Steinmetz et al, 2019;Wekselblatt et al, 2016). However, they are still far from being whole-brain and multi-area simultaneous access is still limited.…”

Section: Introductionmentioning

confidence: 99%

Functional MRI of large scale activity in behaving mice

Fonseca

Bergomi

Mainen

et al. 2020

Preprint

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Behaviour involves complex dynamic interactions across many brain regions.Detecting whole-brain activity in mice performing sophisticated behavioural tasks could facilitate insights into distributed processing underlying behaviour, guide local targeting, and help bridge the disparate spatial scales between rodent and human studies. Here, we present a comprehensive approach for recording brain-wide activity with functional magnetic resonance imaging (fMRI) compatible with a wide range of behavioural paradigms and neuroscience questions. We introduce hardware and procedural advances to allow multi-sensory, multi-action behavioural paradigms in the scanner. We identify signal artifacts arising from task-related body movements and propose novel strategies to suppress them. We validate and explore our approach in a 4-odour classical conditioning and a visually-guided operant task, illustrating how it can be used to extract information insofar intangible to rodent behaviour studies.Our work paves the way for future studies combining fMRI and local circuit techniques during complex behaviour to tackle multi-scale behavioural neuroscience questions.

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“…In other words, the volumetric information is projected into a single 2D image. As such, extended depth of focus (EDF) imaging can be attractive to image sparsely populated structures rapidly and has found promising applications in functional imaging of neuronal activity [1,2].…”

Section: Introductionmentioning

confidence: 99%

Extended depth of focus multiphoton microscopy via incoherent pulse splitting

Chen

Chakraborty

Daetwyler

et al. 2020

Preprint

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We present a phase mask that can be easily added to any multi-photon raster scanning microscope to extend the depth of focus five-fold at a small loss in lateral resolution. The method is designed for ultrafast laser pulses or other light-sources featuring a low coherence length. In contrast to other methods of focus extension, our approach uniquely combines low complexity, high light-throughput and multicolor capability. We characterize the point-spread function in a two-photon microscope and demonstrate fluorescence imaging of GFP labeled neurons in fixed brain samples as imaged with conventional and extended depth of focus two-photon microscopy.

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Rapid mesoscale volumetric imaging of neural activity with synaptic resolution

Cited by 104 publications

References 28 publications

Improving laser standards for three-photon microscopy

Improving laser standards for three-photon microscopy

Functional MRI of large scale activity in behaving mice

Extended depth of focus multiphoton microscopy via incoherent pulse splitting

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